Marine benthic nitrogen (N) cycling may vary widely across space and seasons; it is thus needed to make high-resolution estimations of these important ecosystem processes with a reasonable number of variables. In this study, we determined the benthic denitrification and anammox potentials in two basins, the Bohai Sea and the North Yellow Sea of China in May and November, and evaluated models in predicting these functions with environmental factors and/or microbial gene-based functional traits. We found that denitrification generally dominated the N loss (54-98%), and that the denitrification rate varied greatly between basins and seasons. The anammox rate was generally higher in the Bohai Sea than in the North Yellow Sea in both seasons, and it made a greater contribution in November (22%) than in May (16%). Among the measured environmental factors, chlorophyll a in bottom waters and sedimentary organic carbon content were the most influential for predicting denitrification and anammox rates, respectively. On the other hand, the alpha diversities and gene abundances of involved bacteria were poorly correlated with the function potentials, indicating that these functional traits could not well explain the functions alone. Upon the incorporation of two gene copy number ratios [nosZ/(nirS + nirK) and nirK/bacterial 16S rRNA genes] into the environmental factor-parameterized models, however, we found that the predictive powers of the regression models for total N loss, denitrification and anammox rates, and contributions of anammox increased substantially, indicating that taking microbial functional traits into account could make estimations of these N-cycling functions in coastal ecosystems more accurate. contributed equally to this work. Key Points:• Benthic N 2 productions via denitrification and anammox varied greatly across seasons and at a basin-wide scale • Bottom water chlorophyll a and sediment organic carbon were the main environmental factors underlying the biogeography of benthic N loss • Adding proper functional traits into models improved the predictability for benthic denitrification and anammox functionsSupporting Information:• Supporting Information S1

The spatial and temporal patterns of diversity, community structure, and their drivers are fundamental issues in microbial ecology. This study aimed to investigate the relative importance of spatial and seasonal controls on the distribution of nitrogen cycling microbes in sediments of estuarine tidal flats, and to test the hypothesis that metals impact the distribution of nitrogen-cycling microbes in the coastal system. Two layers of sediment samples were collected from three estuarine tidal flats of Laizhou Bay in 2010 winter and 2011 summer. The alpha diversities (Shannon and Simpson indices) and community structure of ammonia oxidizing bacteria (AOB) and archaea (AOA), denitrifier and anammox bacteria (AMB) were revealed using denaturing gradient gel electrophoresis and clone library analysis of amoA, nosZ and 16S rRNA gene markers. We found that both AOB and AMB exhibited distinct seasonal patterns in either alpha diversity or community turnover; AOA had different alpha diversities in two layers, but neither spatial nor seasonal patterns were found for their community turnover. However, no distinct spatiotemporal pattern was observed for either diversity or community composition of nosZ-type denitrifiers. For correlations between alpha diversities and environmental factors, significant correlations were found between AOB and ammonium, temperature and As, between denitrifiers and nitrite, salinity and Pb, and between AMB and Pb, ratio of organic carbon to nitrogen, ammonium, pH and dissolved oxygen. Salinity and sediment grain size were the most important factors shaping AOB and AOA communities, respectively; whereas AMB community structure was mostly determined by temperature, dissolved oxygen, pH and heavy metals As and Cd. These results stress that ammonia oxidizers, denitrifiers and anammox bacteria have generally different distributional patterns across time and space, and heavy metals might have contributed to their differentiated distributions in coastal sediments.

Denitrifi cation and anammox processes are major nitrogen removal processes in coastal ecosystems. However, the spatiotemporal dynamics and driving factors of the diversity and community structure of involved functional bacteria have not been well illustrated in coastal environments, especially in human-dominated ecosystems. In this study, we investigated the distributions of denitrifi ers and anammox bacteria in the eutrophic Bohai Sea and the northern Yellow Sea of China in May and November of 2012 by constructing clone libraries employing nosZ and 16S rRNA gene biomarkers. The diversity of nosZ-denitrifi er was much higher at the coastal sites compared with the central sites, but not signifi cant among basins or seasons. Alphaproteobacteria were predominant and prevalent in the sediments, whereas Betaproteobacteria primarily occurred at the site near the Huanghe (Yellow) River estuary. Anammox bacteria Candidatus Scalindua was predominant in the sediments, and besides, Candidatus Brocadia and Candidatus Kuenenia were also detected at the site near the Huanghe River estuary that received strong riverine and anthropogenic impacts. Salinity was the most important in structuring communities of nosZdenitrifi er and anammox bacteria. Additionally, anthropogenic perturbations (e.g. nitrogen overloading and consequent high primary productivity, and heavy metal discharges) contributed signifi cantly to shaping community structures of denitrifi er and anammox bacteria, suggesting that anthropogenic activities would infl uence and even change the ecological function of coastal ecosystems.